6-Mercapto-cyclodextrin derivatives: reversal agents for drug-induced neuromuscular block

ABSTRACT

Disclosed is a 6-mercapto-cyclodextrin derivative having a general formula (I, ) wherein m is 0-7 and n is 1-8 and m+n=7 or 8; R is (C 1-6 ) alkylene, optionally substituted with 1-3 OH groups, or (CH 2 ) o -phenylene-CH 2 ) p —; o and p are independently 0-4; X is COOH, CONHR 3 , NHCOR 2 , SO 2 OH, PO(OH) 2 , O(CH 2 —CH 2 —O) q —H, OH or tetrazol-5-yl: R 2  is H or (C 1-3 )alkyl; R 2  is carboxyphenyl; q is 1-3; or pharmaceutically acceptable salts thereof. The 6-mercaptocyclodextrin derivative is highly suitable for use in the reversal of drug-induced neuromuscular block.

RELATED APPLICATION

This application is a divisional of U.S. application Ser. No.10/148,307, filed May 29, 2002, now U.S. Pat. No. 6,670,340, which is a371 of PCT/EP00/11789, filed Nov. 23, 2000.

The invention relates to 6-mercapto-cyclodextrin derivatives, to theiruse for the preparation of a medicament for the reversal of drug-inducedneuromuscular block, and to a kit for providing neuromuscular block andits reversal.

A neuromuscular blocking agent (NMBA, also called a muscle relaxant) isroutinely used during the administration of anaesthesia to facilitateendotracheal intubation and to allow surgical access to body cavities,in particular the abdomen and thorax, without hindrance from voluntaryor reflex muscle movement. NMBAs are also used in the care ofcritically-ill patients undergoing intensive therapy, to facilitatecompliance with mechanical ventilation when sedation and analgesia alonehave proved inadequate, and to prevent the violent muscle movements thatare associated with electroconvulsive therapy treatment.

Based on their mechanisms of action, NMBAs are divided into twocategories: depolarizing and non-depolarizing. Depolarizingneuromuscular blocking agents bind to nicotinic acetylcholine receptors(nAChRs) at the neuromuscular junction in a way similar to that of theendogenous neurotransmitter acetylcholine. They stimulate an initialopening of the ion channel, producing contractions known asfasciculations. However, since these drugs are broken down onlyrelatively slowly by cholinesterase enzymes, compared to the very rapidhydrolysis of acetylcholine by acetylcholinesterases, they bind for amuch longer period than acetylcholine, causing persistent depolarizationof the end-plate and hence a neuromuscular block. Succinylcholine(suxamethonium) is the best known example of a depolarizing NMBA.

Non-depolarizing neuromuscular blocking agents compete withacetylcholine for binding to muscle nAChRs, but unlike depolarizingNMBAs, they do not activate the channel. They block the activation ofthe channel by acetylcholine and hence prevent cell membranedepolarization, and as a result, the muscle will become flaccid. Most ofthe clinically-used NMBAs belong to the non-depolarizing category. Theseinclude tubocurarine, atracurium, (cis)atracurium, mivacurium,pancuronium, vecuronium, rocuronium and rapacuronium (Org 9487).

At the end of surgery or a period of intensive care, a reversal agent ofNMBAs is often given to the patient to assist the recovery of musclefunction. Most commonly used reversal agents are inhibitors ofacetylcholinesterase (AChE), such as neostigmine, edrophonium andpyridostigmine. Because the mechanism of action of these drugs is toincrease the level of acetylcholine at the neuromuscular junction byinhibiting the breakdown of acetylcholine, they are not suitable forreversal of depolarizing NMBAs such as succinylcholine. The use of AChEinhibitors as reversal agents leads to problems with selectivity, sinceneurotransmission to all synapses (both somatic and autonomic) involvingthe neurotransmitter acetylcholine is potentiated by these agents. Thisnon-selectivity may lead to many side-effects due to the non-selectiveactivation of muscarinic and nicotinic acetylcholine receptors,including bradycardia, hypotension, increased salivation, nausea,vomiting, abdominal cramps, diarrhoea and bronchoconstriction. Thereforein practice, these agents can be used only after or together with theadministration of atropine (or glycopyrrolate) to antagonize themuscarinic effects of acetylcholine at the muscarinic receptors in theautonomic parasympathetic neuro-effector junctions (e.g. the heart). Theuse of a muscarinic acetylcholine receptor (mAChR) antagonist such asatropine causes a number of side-effects, e.g., tachycardia, dry mouth,blurred vision, difficulties in emptying the bladder and furthermore mayaffect cardiac conduction.

A further problem with anticholinesterase agents is that residualneuro-muscular activity must be present (>10% twitch activity) to allowthe rapid recovery of neuromuscular function. Occasionally, either dueto hyper-sensitivity of the patient or accidental overdose,administration of NMBAs can cause complete and prolonged block ofneuromuscular function (“profound block”). At present, there is noreliable treatment to reverse such a ‘profound block’. Attempts toovercome a ‘profound block’ with high doses of AChE inhibitors has therisk of inducing a “cholinergic crisis”, resulting in a broad range ofsymptoms related to enhanced stimulation of nicotinic and muscarinicreceptors.

In European Patent Application 99,306,411 (AKZO NOBEL N.V.) the use ofchemical chelators (or sequestrants) as reversal agents has beendisclosed. Chemical chelators capable of forming a guest-host complexfor the manufacture of a medicament for the reversal of drug-inducedneuromuscular block were described. The use of chemical chelators asreversal agents for NMBAs has the advantage that they are effective inreversing the action of both depolarizing and non-depolarizing NMBAs.Their use does not increase the level of acetylcholine and thereforethey produce fewer side effects and none associated with the stimulationof muscarinic and nicotinic receptors seen with the AChE reversalagents. In addition, there is no need for the combined use of an AChEinhibitor and a mAChR antagonist (e.g., atropine), while the chemicalchelators may further be safely employed for the reversal of ‘profoundblock’. Examples of such chemical chelators, as disclosed in EP99,306,411, were selected from various classes of, mostly cyclic,organic compounds which are known for their ability to form inclusioncomplexes with various organic compounds in aqueous solution, e.g.cyclic oligosaccharides, cyclophanes, cyclic peptides, calixarenes,crown ethers and aza crown ethers.

The cyclodextrins,

a class of cyclic molecules containing six or more α-D-glucopyranoseunits linked at the 1,4 positions by α-linkages as in amylose, andderivatives thereof, were identified in EP 99306411 as particularlyuseful in the reversal of many of the commonly used neuromuscularblocking agents, or muscle relaxants, such as rocuronium, pancuronium,vecuronium, rapacuronium, mivacurium, atracurium, (cis)atracurium,succinylcholine and tubocurarine,

It has now been found that 6-mercapto-cyclodextrin derivatives havingthe general formula I

wherein m is 0-7 and n is 1-8 and m+n=7 or 8;

-   R is (C₁₋₆)alkylene, optionally substituted with 1-3 OH groups, or    (CH₂)_(o)-phenylene-(CH₂)_(p)—;-   o and p are independently 0-4;-   X is COOH, CONHR₁, NHCOR₂, SO₂OH, PO(OH)₂, O(CH₂—CH₂—O)_(q)—H, OH or    tetrazol-5-yl;-   R₁ is H or (C₁₋₃)alkyl;-   R₂ is carboxyphenyl;-   q is 1-3;-   or pharmaceutically acceptable salts thereof;-   are highly active in vivo in the reversal of the action of    neuromuscular blocking agents.

No protection per se is sought for the following 6-mercapto-cyclodextrinderivatives:

-   6-per-deoxy-6-per-(2-hydroxyethylthio)-β-cyclodextrin and-   6-per-deoxy-6-per-(2-hydroxyethylthio)-γ-cyclodextrin, which are    described by Ling, C. and Darcy, R. (J. Chem. Soc. Chem Comm. 1993,    (2), 203-205);-   6-mono-deoxy-6-mono-(2-hydroxyethylthio)-β-cyclodextrin, which is    disclosed by Fujita, K. et al. (Tetr. Letters 21, 1541-1544, 1980);-   6-per-deoxy-6-per-(carboxymethylthio)-β-cyclodextrin, which is    described by Guillo, F. et al. (Bull. Chem. Soc. Chim. Fr. 132 (8),    857-866, 1995);-   6-mono-deoxy-6-mono-(carboxymethylthio)-β-cyclodextrin, which is    described by Akiie, T. et al. (Chem. Lett. 1994 (6), 1089-1092);-   6A,6B-dideoxy-6A,6B-bis[(o-carboxyphenyl)thio]-p-cyclodextrin and-   6A,6B-dideoxy-6A,6B-bis(carboxymethylthiol)-β-cyclodextrin, which    are described by Tubashi, I. et al. (J. Am. Chem. Soc. 108,    4514-4518, 1986; and-   6-per-deoxy-6-per-(2,3-dihydroxypropylthio)-β-cyclodextrin, which is    described by Baer, H. H. and Santoyo-González, F. (Carb. Res. 280,    315-321, 1996). These prior art 6-mercapto-cyclodextrin derivatives    have been described in relation with different utilities in each    instance.

However, the above mentioned prior art 6-mercapto-cyclodextrinderivatives do belong to the main aspect of the present invention whichrelates to the use of a 6-mercapto-cyclodextrin derivative according tothe general formula I for the manufacture of a medicament for thereversal of drug-induced neuromuscular block.

In one embodiment the invention relates to 6-mercapto-cyclodextrinderivatives having the general formula I,

-   wherein m is 0-7 and n is 1-8 and m+n=7 or 8;-   X is COOH, OH or CONHCH₃;-   R is (C₁₋₆)alkylene or (CH₂)_(o)-phenylene-(CH₂)_(p);-   o and p are independently 0-4; or a pharmaceutically acceptable salt    thereof, with the exclusion of-   6-per-deoxy-6-per-(2-hydroxyethylthio)-β-cyclodextrin;-   6-mono-deoxy-6-mono-(2-hydroxyethylthio)-cyclodextrin;-   6-per-deoxy-6-per-(2-hydroxyethylthio)-γ-cyclodextrin;-   6-per-deoxy-6-per-(carboxymethylthio)-β-cyclodextrin;-   6-mono-deoxy-6-mono-(carboxymethylthio)-β-cyclodextrin;-   6A,6B-dideoxy-6A,6B-bis[(o-carboxyphenyl)thio]-β-cyclodextrin; and-   6A,6B-dideoxy-6A,6B-bis(carboxymethylthiol)-β-cyclodextrin.

The term (C₁₋₆)alkylene as used in the definition of formula I means abranched or straight chain bivalent carbon radical containing 1-6 carbonatoms, such as methylene, ethylene (1,2-ethandiyl), propylene(1-methyl-1,2-ethanediyl), 2-methyl-1,2-ethanediyl,2,2-dimethyl-1,2-ethanediyl, 1,3-propanediyl, 1,4-butanediyl,1,5-pentanediyl and 1,6-hexanediyl.

The term phenylene means a bivalent moiety the free valencies of whichcan be positioned either ortho, meta or para to one another.

The term (C₁₋₃)alkyl means a branched or straight chain alkyl groupcontaining 1-3 carbon atoms, i.e. methyl, ethyl, propyl and isopropyl.

The term carboxyphenyl means a phenyl group which is substituted ateither the ortho-, the meta- or the para-position with a carboxy-group.The ortho-carboxyphenyl group is preferred.

Compounds according to formula I wherein n+m is 7 are derivatives ofβ-cyclodextrin, those wherein n+m is 8 are derived from γ-cyclodextrin.

Preferred are the 6-mercapto-cyclodextrin derivatives of formula Iwherein X is COOH, or a pharmaceutically acceptable salt thereof.

More preferred are the 6-mercapto-γ-cyclodextrin derivatives of formulaI wherein n is 8, R is (C₁₋₆)alkylene and X is COOH.

Particularly preferred 6-mercapto-cyclodextrin derivatives of theinvention are

-   6-per-deoxy-6-per-(2-carboxyethyl)thio-γ-cyclodextrin;-   6-per-deoxy-6-per-(3-carboxypropyl)thio-γ-cyclodextrin;-   6-per-deoxy-6-per-(4-carboxyphenyl)thio-γ-cyclodextrin;-   6-per-deoxy-6-per-(4-carboxyphenyl methyl)thio-γ-cyclodextrin;-   6-per-deoxy-6-per-(2-carboxypropyl)thio-γ-cyclodextrin; and-   6-per-deoxy-6-per-(2-sulfoethyl)thio-γ-cyclodextrin.

The 6-mercapto-cyclodextrin derivatives of formula I can be prepared byreacting a C6-activated cyclodextrin derivative of formula II with analkylthiol, arylalkylthiol or arylthiol derivative corresponding toH—S—R—X, wherein R and X have the meaning as previously defined, in thepresence of an inorganic or organic base.

Formula II wherein m is 0-7, n is 1-8, m+n=7 or 8 and Y is a leavinggroup which can be a halide (Cl, Br or I), sulfuric ester or a sulfonicester function, such as a tosylate, a napthtalenesulfonate or atriflate.

Conversely the 6-mercapto-cyclodextrin derivatives of formula I can alsobe prepared by reacting a 6-thiol γ- or β-cyclodextrin derivative offormula III with an alkylating agent, e.g., alkyl halide, arylalkylhalide, alkyl sulfonate, arylalkyl sulfonate, corresponding to Y—X—R,wherein Y, X and R have the meanings as previously defined, or with adouble bond containing reagent, e.g., vinyl alkane, acrylate, etc., oran epoxide in the presence of an inorganic or organic base.

Formula III wherein m is 0-7, n is 1-8, m+n=7 or 8.

Alternative synthesis routes for the preparation of the6-mercapto-cyclodextrin derivatives of the invention are known to theskilled person. The chemistry of the derivatisation of cyclodextrins iswell documented (see for example: Comprehensive SupramolecularChemistry, Volumes 1-11, Atwood J. L., Davies J. E. D., MacNicol D. D.,Vogtle F., eds; Elsevier Science Ltd., Oxford, UK, 1996).

Pharmaceutically acceptable salts of 6-mercapto-cyclodextrin derivativesof formula I wherein X represents the carboxylic acid group COOH, thesulphonic acid group SO₂OH, the phosphonic acid group PO(OH)₂ or thetetrazol-5-yl group, may be obtained by treating the acid with anorganic base or a mineral base, like sodium-, potassium- or lithiumhydroxide.

The 6-mercapto-cyclodextrin derivatives, or pharmaceutically acceptablesalts or solvates thereof, for use in the invention are administeredparenterally. The injection route can be intravenous, subcutaneous,intradermal, intramuscular, or intra-arterial. The intravenous route isthe preferred one. The exact dose to be used will necessarily bedependent upon the needs of the individual subject to whom themedicament is being administered, the degree of muscular activity to berestored and the judgement of the anaesthetist/critical-care specialist.Extracorporal application of the chemical chelators of the invention,for instance by mixing of the chemical chelator with the blood duringdialysis or during plasmapheresis, is also contemplated.

In a further aspect the invention relates to a kit for providingneuromuscular block and its reversal comprising (a) a neuromuscularblocking agent, and (b) a 6-mercapto-cyclodextrin derivative accordingto general formula I capable of forming a guest-host complex with theneuromuscular blocking agent. With a kit according to the invention ismeant a formulation, which contains separate pharmaceuticalpreparations, i.e. the neuromuscular blocking agent and a6-mercapto-cyclodextrin derivative of formula I, i.e. the reversalagent. The components of such a kit of parts are to be usedsequentially, i.e. the neuromuscular blocking agent is administered to asubject in need thereof, which is followed, at a point in time whenrestoration of muscle function is required, by the administration of thereversal agent, i.e. a 6-mercapto-cyclodextrin derivative of the presentinvention.

A preferred kit, according to the invention, contains a6-mercapto-cyclodextrin derivative of formula I and a neuromuscularblocking agent which is selected from the group consisting ofrocuronium, vecuronium, pancuronium, rapacuronium, mivacurium,atracurium, (cis)atracurium, tubocurarine and suxamethonium. Aparticularly preferred kit of the invention comprises rocuronium as theneuromuscular blocking agent.

Mixed with pharmaceutically suitable auxiliaries and pharmaceuticallysuitable liquids, e.g. as described in the standard reference, Gennaroet al., Remington's Pharmaceutical Sciences, (18th ed., Mack PublishingCompany, 1990, Part 8: Pharmaceutical Preparations and TheirManufacture; see especially Chapter 84 on “Parenteral preparations, pp.1545-1569; and Chapter 85 on “Intravenous admixtures”, pp. 1570-1580)the 6-mercapto-cyclodextrin derivatives can be applied in the form of asolution, e.g. for use as an injection preparation.

Alternatively, the pharmaceutical composition may be presented inunit-dose or multi-dose containers, for example sealed vials andampoules, and may be stored in a freeze dried (lyophilised) conditionrequiring only the addition of the sterile liquid carrier, for example,water prior to use.

The invention further includes a pharmaceutical formulation, ashereinbefore described, in combination With packaging material suitablefor said composition, said packaging material including instructions forthe use of the composition for the use as hereinbefore described.

The invention is illustrated in the following examples.

EXAMPLE 1 6-Mono-deoxy-6-mono-(4-carboxyphenyl)thio-γ-cyclodextrin,sodium salt

To a round bottom flask containing pyridine (120 ml) was added dryγ-cyclodextrin (2.0 g, 1.54 mmol) under nitrogen at room temperature.After dissolution, 2-napthalenesulfonyl chloride (1.05 g, 4.64 mmol) inpyridine (20 ml) was added and the mixture stirred for 24 h. Quenchedwith water (50 ml) and evaporated to dryness to leave crude6-mono-O-(2′-naphthalenesulfonyl)-γ-cyclodextrin.

Sodium hydride (0.38 g, 15.83 mmol) was suspended in drydimethylformamide (20 ml). 4-Mercaptobenzoic acid (0.7 g, 4.55 mmol) wasthen added to the suspension and the resulting mixture was stirred for20 minutes. γ-Cyclodextrin nosylate (3.2 g, 2.12 mmol) was added to themixture and the reaction was heated to 100° C. for 90 minutes. Aftercooling, acetone was added to precipitate a solid, which wasreprecipitated from water/acetone. This was then dissolved in water (20ml), pH adjusted to 7.0 by adding 2N hydrochloric acid, thenchromatographed on a Sephadex DEAE A-25 column. Appropriate fractionswere combined, dialysed, then precipitated, twice from water/acetone togive 400 mg of the titled compound. ¹H NMR in DMSO δ 7.4 to 7.8 (ArH),5.0 to 5.2 (8 H), 4.13 (1 H), 3.7 to 4.0 (29 H), 3.7 to 3.4 (17 H), 3.25(1 H) ppm. ¹³C NMR in DMSO δ 129.9 an 127.5 (ArC), 103.3 and 102.9 (C1and C1′), 85.0 (C4′), 81.6 (C4), 73.8 (C3(C2), 72.2 (C5), 70.8(C5′),60.6 (C6), 34.3 (C6′) ppm. Electrospray MS [M+1455.7 and [M+Na]⁺=1477.7.

EXAMPLE 2 6-Mono-deoxy-6-mono-(2-carboxylphenyl)thio-γ-cyclodextrin,sodium salt

Sodium hydride (60% dispersed in oil, 0.18 g, 4.5 mmol) was added tothiosalicylic acid (0.34 g, 2.2 mmol) in DMF (25 ml) in one portion andstirred at room temperature for 30 min. To this was then added the crudesolution of 6-mono-O-(2′-naphthalenesulfonyl)-γ-cyclodextrin (2.5 g,1.45 mmol) in DMF (15 ml) and heated to 70° C. for 24 h. The mixture wascooled and quenched with water (20 ml) before evaporating to dryness.Water was then added to the residue and the resulting solution waspoured into acetone (250 ml) to effect precipitation. The resultingsolid was collected by filtration and dissolved in water (10 ml) beforepassing through a Sephadex DEAE A-25 column eluting with water then 0.2N NaOH. Fractions containing the product were combined and evaporated toa low volume and dialysed (MWCO 1000) by changing the external waterfour times. Internal solution was evaporated to low volume and pouredinto acetone (100 ml). Solid was collected by filtration and dried undervacuum at 70° C. to leave the title compound (235 mg) as a white solid.¹H NMR (D₂O) δ 7.50-7.10 (4H, m, Ar—H), 5.14 (8 H, m, CyD 1-H), 4.16(1H, m, CyD 5-H), 3.98-3.85 (26 H, m, CyD 3,5,2,4,-H), 3.70-3.61 (20 H,m, CyD 2,3,4,6-H), 3.15 (1 H, m, CyD 6-H) ppm; Electrospray MS m/z1477.6 for [M+Na]⁺, calcd for C₅₅H₈₃NaO₄₁S M 1455.304.

EXAMPLE 3 6-per-deoxy-6-per-(3-carboxyphenyl)thio-γ-cyclodextrin, sodiumsalt

Triphenylphosphine (30.1 g, 15 eq) was dissolved with stirring in dryDMF (160 ml). To this was added iodine (30.5 g, 15.6 eq) over 10 min.with heat evolved. Dry γ-cyclodextrin (10 g, 7.7 mmol) was then addedand the mixture was heated to 70° C. for 24 h. The mixture was allowedto cool, to which sodium methoxide (3.1 g sodium in 50 ml methanol) wasadded and the mixture was stirred for 30 min, before pouring ontomethanol (800 ml) and evaporating to dryness. To the residue was addedwater (500 ml) and the solid was collected by filtration and washed withwater (3×100 ml), then acetone (3×100 ml), and dried under vacuum at 70°C. to give 6-per-deoxy-6-per-iodo-γ-cyclodextrin as a yellow solid (16.2g) which was used without further purification.

To a solution of 3-mercaptobenzoic acid (1.0 g, 10 eq) in DMF (30 ml)was added 60% sodium hydride dispersed in oil (476 mg, 22 eq)portionwise over 30 min. The mixture was cooled and6-per-deoxy-6-per-iodo-γ-cyclodextrin (1.4 g) in DMF (30 ml) was added.The mixture was then stirred at 70° C. for 24 h. The mixture was allowedto cool to room temperature and quenched with the addition of water (20ml) before evaporating to a low volume. The solution was poured intoacetone (500 ml) and the precipitate was collected by filtration,dissolved in water (20 ml) and dialysed (MWCO 1000) by changing theexternal water four times. Internal solution was evaporated to lowvolume and poured into acetone (250 ml). The solid precipitate wascollected by filtration and dried under vacuum at 70° C. to afford thetitle compound (1.45 g) as a white solid: ¹H NMR (D₂O) δ 7.77 (8H, br s,Ar—H), 7.55 (8H, d, J 6.0 Hz, Ar—H), 7.71 (16H, m, Ar—H), 5.16 (8H, s,CyD 1-H), 4.00-3.94 (16H, m, CyD 3,5-H), 3.58-3.53 (16H, m, CyD 4,2-H),3.43-3.40 (8H, m, CyD 6-H), 3.24-3.20 (8H, m, CyD 6-H); Electrospray m/z1190.6 for [M-8Na+6H]²⁻, calcd for C₁₀₄H₁₀₄Na₈O₄₈S₈ M 2562.39.

EXAMPLE 4 6-Per-deoxy-6-per-(2-carboxyethyl)thio-γ-cyclodextrin, sodiumsalt

3-Mercaptopropionic acid (1.22 ml, 14.0 mmol) was dissolved in dry DMF(45 ml) under N₂ at room temperature. To this solution was added inthree portions sodium hydride (1.23 g, 30.8 mmol, 60%) and the mixturewas stirred for a further 30 min. To this mixture was then addeddropwise a solution of 6-per-deoxy-6-per-iodo-γ-cyclodextrin (3.12 g,1.40 mmol) in 45 ml dry DMF. After addition, the reaction mixture washeated at 70° C. for 12 h. After cooling, water (10 ml) was added to themixture and the volume was reduced to 40 ml in vacuo, to which ethanol(250 ml) was added resulting in precipitation. The solid precipitate wascollected by filtration and dialysed for 36 h. The volume was thenreduced to 20 ml in vacuo. To this was added ethanol, and theprecipitate was collected by filtration and dried to give the titlecompound as a white solid (1.3 g, 43%). ¹H-NMR D₂O δ 2.47-2.51 (m, 16H);2.84-2.88 (m, 16H); 3.00-3.02 (t, 8H); 3.11-3.14 (t, 8H); 3.62-3.68 (m,16H); 3.92-3.97 (m, 8H); 4.04-4.06 (m, 8H); 5.19 (m, 8H) ppm. MS FIA+ion at 2024.9 m/z.

EXAMPLE 5 6-Per-deoxy-6-per-(5-carboxypentyl)thio-γ-cyclodextrin, sodiumsalt

The title compound was prepared in a similar way as described forExample 4 by reacting 6-mercaptohexanoic acid (1.34 g, 0.90 mmol) with6-per-deoxy-6-per-iodo-γ-cyclodextrin. ¹H-NMR D₂O δ 1.40 (s, 16H);1.57-1.64 (m, 32H); 2.17-2.21 (m, 16H); 2.67-3.00 (m, 16H); 2.85-2.90(m, 8H); 3.15-3.20 (m, 8H); 3.52-3.59 (m, 8H); 3.60-3.63 (m, 8H);3.87-3.93 (m, 16H); 5.16 (s, 8H) ppm. MS FIA +ions at 2362.2, 2213, 2065and 1919 m/z

EXAMPLE 6 6-Per-deoxy-6-per-(3-carboxypropyl)thio-γ-cyclodextrin, sodiumsalt

The title compound was prepared in a similar way as described forExample 4 by reacting 4-mercaptobutyric acid (1.10 g, 0.009 mol) with6-per-deoxy-6-per-iodo-γ-cyclodextrin. ¹H-NMR D₂O δ 1.87-1.88 (m, 16H);2.27-2.30 (m, 16H); 2.67-2.71 (m 16H); 2.98-3.00 (m, 8H); 3.13-3.16 (m,8H); 3.61-3.63 (m, 16H); 3.94-4.03 (m 16H); 5.21 (s, 8H) ppm. MS FIA+ion at 2138.8 m/z.

EXAMPLE 7 6-Per-deoxy-6-per-carboxymethylthio-γ-cyclodextrin, sodiumsalt

Sodium hydride (60% dispersion, 0.34 g, 8.60 mmol) was added to astirred solution of ethyl 2-mercaptoacetate (0.92 ml, 8.40 mmol) in DMF(20 ml) under nitrogen at room temperature. After effervescence hadceased (15 min), per-6-deoxy-per-6-iodo-γ-cyclodextrin (2.17 g, 1.00mmol) was added to the system. After a further 5 min, the temperaturewas raised to 70° C. and the reaction was left with stirring for 17 h.After cooling, DMF was removed in vacuo. Methanol (50 ml) was added anda creamy white solid slowly crystallised out of solution. This wasfiltered off under suction, washed with methanol and dried to give6-per-deoxy-6-per-carbethoxymethylthio-γ-cyclodextrin as a solid (1.74g, 82%). δ_(H)(d6-dmso) 4.95-4.85 (8H, m, 8×anomeric CH), 4.05 (16H, q,8×CH ₂CH₃), 3.85-3.75 (8H, m), 3.60-3.50 (8H, m), 3.40-3.20 (32H, bs,8×CH₂SCH₂), 3.20-3.10 (8H, m), 2.95-2.85 (8H, m), 1.20 (24H, t, 8×CH₂CH₃).

To 1 M solution of sodium hydroxide (7 ml) was added6-per-deoxy-6-per-carbethoxymethylthio-γ-cyclodextrin (1.00 g, 0.47mmol) and the reaction was allowed to stir at room temperature. After18h, the clear solution was dialysed for 8 h, with water (2 L) beingreplaced every 2 h. After this time, the contents of the dialysis tubingwas emptied into a flask and water evaporated in vacuo, giving the titlecompound as a white solid (0.62 g, 64%). δ_(H)(D₂O) 5.21 (8H, d,8×anomeric CH), 4.18-4.05 (8H, m), 4.00 (8H, dd), 3.78 (8H, dd), 3.70(8H, dd), 3.40 (16H, dd), 3.20 (8H, d), 3.02 (8H, dd). δ_(C)(D₂O) 178.1,101.6, 82.8, 73.0, 72.7, 71.8, 39.0, 34.1 LC/MS TOF 1889 m/z

EXAMPLE 8 6-Per-deoxy-6-per-(4-carboxyphenyl)thio-γ-cyclodextrin, sodiumsalt

To a solution of 4-mercaptobenzoic acid (856 mg) in DMF (30 ml) wasadded 60% sodium hydride dispersed in oil (372 mg) portionwise over 30min. The mixture was cooled and per-6-deoxy-per-6-bromo-γ-cyclodextrin(1.0 g) was added in one portion and the mixture was stirred at 70° C.for 24 h. The mixture was allowed to cool to room temperature andquenched with the addition of water (20 ml) before evaporating to a lowvolume. The solution was poured into ethanol (250 ml) and theprecipitate was; collected by filtration, dissolved in water (20 ml) anddialysed (MWCO 1000) by changing the external water four times. Internalsolution was evaporated to low volume and poured into acetone (250 ml).The solid precipitate was collected by filtration and dried under vacuumat 70° C. to afford the title compound (1.2 g) as a white solid. ¹H NMR(D₂O, 343 K) δ 7.70 (16H, d, J=8.1 Hz, Ar—H), 7.23 (16H, d, J=7.3 Hz,Ar—H), 5.15 (8H, s, CyD 1-H), 4.00-3.96 (16H, m, CyD 3,5-H), 3.55-3.53(24H, m, CyD 6′,4,2-H), 3.15 (8H, m, CyD 6-H); MALDI-TOF m/z 2383.7 for[M−Na₈+H₆], calcd for C₁₀₄H₁₀₄Na₈O₄₈S₈ M 2562.39.

EXAMPLE 9 6-Per-deoxy-6-per-(4-carboxymethylphenyl)thio-γ-cyclodextrin,sodium salt

To a solution of 4-mercaptophenylacetic acid (10 eq) in DMF (50 ml) wasadded 60% sodium hydride in oil (22 eq) portionwise over 30 min. Themixture was cooled and per-6-deoxy-per-6-bromo-γ-cyclodextrin (1.0 g)was added in one portion and the mixture was stirred at 70° C. for 24 h.The mixture was allowed to cool to room temperature and quenched withthe addition of water (20 ml) before evaporating to a low volume. Thesolution was then poured into acetone (250 ml) and the precipitate wascollected by filtration, suspended in water (20 ml) and dialysed (MWCO1000) by changing the external water four times. Internal solution wasevaporated to low volume and poured into acetone (250 ml). The solidprecipitate was collected by filtration and dried under vacuum at 70° C.to afford the title compound (1.44 g) as a white solid. ¹H NMR (D₂O,343K) δ 7.15 (16 H, d, J=8.0 Hz, Ar—H), 6.99 (16H, d, J=8.0 Hz, Ar—H),4.98 (8H, s, CyD 1-H), 3.90-3.72 (16H, m, CyD 3,5-H), 3.51-3.43 (16H, m,CyD 4,2-H), 3.28 (24H, m, CH₂—Ar, CyD 6′-H), 3.15-3.10 (1H, m, CyD 6-H);MALDI-TOF m/z 2495.8 for [M−Na₈+H₆], calcd for C₁₁₂H₁₂₀Na₈O₄₈S₈ M2674.6.

EXAMPLE 10 6-Per-deoxy-6-per(3-amidopropyl)thio-γ-cyclodextrin

To a mixture of 6-per-deoxy-6-per-thio-γ-cyclodextrin (500 mg; preparedas described in Example 17) and potassium iodide (5 mg) in DMF (10 ml)was added 4-chlorobutamide (673 mg; Fries et. al. Biochemistry 1975, 14,5233). Caesium carbonate (1.8 g) was added and the reaction mixture washeated to 60° C. overnight. The resulting mixture was poured intoacetone, filtered, washed with ethanol and water and then dried in-vacuo(118 mg; 16.2%). ¹H NMR (DMSO/D₂O) δ 4.9(1H, s), 3.8 (1H, m), 3.6 (1H,m) 3.4 (2H, m), 3.05 (1H, m), 2.85 (1H, m), 2.2 (2H, m), 1.75 (2H, m).Electrospray Mass Spectrum M−H (m/z) 2105.

EXAMPLE 11 6-per-deoxy-6-per(5-hydroxy-3-oxa-pentyl)thio-γ-cyclodextrin

2-(2-Mercaptoethoxy)ethanol (1.4 g, 11.6 mmol) was dissolved in DMF (20ml) and stirring commenced at room temperature under a nitrogenatmosphere. Per-6-bromo-γ-cyclodextrin (2 g, 1.12 mmol) and caesiumcarbonate (3.2 g, 9.86 mmol) were then added and the resultantsuspension stirred at 60° C. overnight under a nitrogen atmosphere.After cooling to room temperature the suspension was poured into acetone(200 ml) and the insoluble material isolated by filtration, washed withacetone (×3) and dried in vacuo. The crude product was dissolved inde-ionised water (20 ml) and dialysed (10 h). The contents of thedialysis membrane were then concentrated in vacuo to yield 1 g of thedesired product as a cream solid.

¹H NMR (D₂O, 400 MHz): δ 2.81.3.00 (m, 24H), 3.21-3.31 (d, 8H), 3.49 (t,8H), 3.55-3.75 (m, 56H), 3.82 (t, 8H), 3.89 (t, 8H), 5.11 (d, 8H).ESI-MS: 2175 (M−H)⁻

EXAMPLE 126-per-deoxy-6-per[(2(2-carboxybenzoyl)amino)ethyl]thio-γ-cyclodextrin,sodium salt

Per-6-mercapto-γ-cyclodextrin (1 g, 0.7 mmol; see example 17) wasdissolved in DMF (10 ml) and stirring commenced at room temperatureunder a nitrogen atmosphere. N-(2-Bromoethyl)phthalimide (1.57 g, 6.17mmol) and caesium carbonate (2 g, 6.17 mmol) were added and theresultant suspension was stirred at 60° C. overnight under a nitrogenatmosphere. After cooling to room temperature the DMF was removed invacuo and water (100 ml) was added with vigorous stirring. Theprecipitate was isolated by filtration, washed with water (×3) and driedin vacuo to yield 1.67 g of a cream solid. Aqueous sodium hydroxide (1M,20 ml) was then added to the crude product (600 mg) and the resultantsolution stirred at room temperature overnight under a nitrogenatmosphere. The solution was then dialysed with de-ionised water untilconstant pH and the contents of the dialysis membrane dried in vacuo toyield 500 mg of the desired product as a glassy solid.

¹H NMR (D₂O, 400 MHz): δ 2.76-2.96 (m, 24H), 3.10-3.30 (m, 8H),3.35-3.62 (m, 32H), 3.78-3.95 (m, 16H), 5.02 (d, 8H), 7.30-7.62 (m,32H); ESI-MS: 1477 (M−2H)²⁻

EXAMPLE 13 6-per-deoxy-6-per(2-hydroxyethyl)thio-γ-cyclodextrin

To a stirred solution of 2-mercaptoethanol (10.85 g, 10 eq) in DMF (500ml) under nitrogen was added 60% sodium hydride dispersed in oil (11.7g, 21 eq) portion-wise over 30 min. The mixture was stirred at roomtemperature for 90 minutes. Per-6-deoxy-6-per-bromo-γ-cyclodextrin (25.0g) was added and the mixture heated to 70° C. for 24 h. The mixture wasallowed to cool to room temperature and quenched by addition of water(50 ml) before evaporating to a low volume. The residue was taken up inwater (100 ml) and poured onto 1:1 methanol/acetone (500 ml). The solidformed was collected by filtration, dissolved in water (500 ml) anddialysed (MWCO 1000) changing the external water four times. Theinternal solution was evaporated to low volume and then re-crystallisedfrom hot water to afford the title compound (8.5 g) as whitecross-shaped crystals.

¹H NMR (400 MHz; DMSO) δ 5.91 (16H, br s, 2,3-OH), 4.92 (8H, s, 1-H),4.71 (8H, t, J 4.4 Hz, SCH₂CH₂OH); 3.75 [8H, t, J 8.0 Hz, 3-H (or 5-H)],3.60-3.50 [24H, m, 5-H (or 3-H), SCH₂CH₂OH], 3.40-3.30 (16H, m, 4-H,2-H), 3.08 (8H, d, J 13.6 Hz, 6-H), 2.82 (8H, dd, J 13.6, 6.8 Hz, 6-H),2.66 (16H, t, J 6.8 Hz, SCH₂CH₂OH); m/z (electrospray) 1775.4 for[M−H]⁻, calcd for C₆₄H₁₁₂S₈O₄₀ M 1776.45. The preparation of thiscompound by a similar method has been published previously: J. Chem.Soc., Chem. Commun., 203 (1993).

EXAMPLE 14 6-per-deoxy-6-per(N-methylamidomethyl)thio-γ-cyclodextrin

To a stirred solution of N-methylmercaptoacetamide (0.58 g, 10 eq) inDMF (30 ml) under nitrogen was added 60% sodium hydride dispersed in oil(0.22 g, 10 eq) portion-wise over 30 min. The mixture was stirred atroom temperature for 30 minutes. Per-6-deoxy-6-per-bromo-γ-cyclodextrin(1.0 g) was added and the mixture heated to 60-70° C. for 48 h. Themixture was allowed to cool to room temperature and quenched by additionof water (20 ml) before evaporating to a low volume. The residualsolution was poured onto ethanol (100 ml). The solid formed wascollected by filtration, dissolved in water (200 ml) and dialysed (MWCO1000), changing the external water four times. The internal solution wasevaporated to low volume and poured onto ethanol (100 ml). Theprecipitate was collected by filtration and dried under vacuum to affordthe title compound (0.55 g) as a white solid.

¹H NMR (400 MHz; D₂O) δ 5.29 (8H, d, J 4.0 Hz, 1-H), 4.10 (8H, br t, J9.6 Hz, 5-H), 4.05 (8H, t, J 9.8 Hz, 3-H), 3.83 (8H, dd, J 10.0, 3.6 Hz,2-H), 3.74 (8H, t, J 9.2 Hz, 4-H), 3.58-3.49 [16H, AB system,SCH₂C(O)NHCH₃], 3.36 (8H, br d, J 12.8 Hz, 6-H), 3.07 (8H, dd, J 14.0,8.4 Hz, 6-H), 2.94 (24H, s, SCH₂C(O)NHCH₃); m/z (electrospray) 1991.7for [M−H]⁻, calculated for C₇₂H₁₂₀N₈S₈O₄₀ M 1992.54.

EXAMPLE 15 6-per-deoxy-6-per(2-carboxypropyl)thio-γ-cyclodextrin, sodiumsalt

Sodium hydride. (60% in oil) (0.44 g) was added to methyl3-mercapto-2-methyl-propionate (1.474 g; J. Med. Chem., 1994, 1159) indimethylformamide (25 ml). After 30 minutesper-6-deoxy-per-6-bromo-γ-cyclodextrin (2.25 g), dissolved indimethylformamide (25 ml), was added. A crystal of sodium iodide wasadded and the mixture heated at 75° C. overnight. The solvent wasdistilled off and the residue crystallised from methanol to give themethyl ester (1.3 g). Mass spec. (M−H) 2224;

¹H NMR (dmso D₆): δ 1.41 (d, 24H), 2.68 (m, 16H), 2.80 (m, 16H), 3.00(m, 8H), 3.61 (3, 24H), 3.79 (m, 8H), 4.95 (s, 8H).

This product was then stirred overnight with sodium hydroxide solution(M, 13 ml). The resulting mixture was, filtered, dialysed to neutrality,and evaporated to dryness to give the title compound (1.13 g). Massspec. (M−H) 2112;

¹H NMR(D₂O): δ 1.15 (d, 24H), 2.5 (m, 8H), 2.65 (m, 8H), 2.8-3.1 (m,24H), 3.65 (m, 16H), 4.0 (m, 16H), 5.2 (s, 8H).

EXAMPLE 16 6-per-deoxy-6-per(3-carboxypropyl)thio-β-cyclodextrin, sodiumsalt

Per-6-deoxy-per-6-bromo-β-cyclodextrin (2.25 g),methyl-4-mercaptobutyrate (1.7 g; Tetrahedron 1998, 2652), cesiumcarbonate (4.24 g) and dimethylformamide (25 ml) were stirred and heatedtogether for three days. The mixture was cooled, poured into water andfiltered. The solid was washed with methanol and dried (2.1 g). This wasstirred overnight with sodium hydroxide solution (M, 21 ml), filteredand the filtrate dialysed to neutrality. This was evaporated to drynessgiving the title compound (1.7 g). Mass Spec.(M−H) 1848.8. ¹H NMR (D₂O):δ 1.75 (m, 16H), 2.15 (m, 16H), 2.6 (m, 16H), 2.85 (m, 8H), 3.05 (m,8H), 3.55 (m, 16H) 3.87 (m, 16H), 5.07 (s, 8H)

EXAMPLE 17 6-Per-deoxy-6-per(2-sulfoethyl)thio-γ-cyclodextrin, sodiumsalt

A: Per-6-deoxy-per-6-thio-γ-cyclodextrin

Per-6-deoxy-per-6-bromo-γ-cyclodextnin (20 g), thiourea (13.5 g) anddimethylformamide (100 ml) were heated together for three days at 65° C.and then ethanolamine (20 ml) was added and heating continued for twohours. The mixture was cooled, diluted with ice water and the productseparated by centrifuge. The solid was washed twice with water and driedin vacuum at 65° C. giving the thiol (7.34 g). Mass spec. (M−H) 1424.

¹H NMR (dmso D₆): δ 2.82 (m, 8H), 3.20 (d, 8H), 3.35 (m, 16H), 6.65 (t,8H), 7.75 ((t, 8H), 5.0 (s, 8H).

B: 6-Per-deoxy-6-per(2-sulfoethyl)thio-γ-clodextrin, sodium salt

The above per-thiol (1 g), 2-bromoethane sulphonic acid sodium salt(1.42 g), cesium carbonate (2.2 g) and dimethylformamide (10 ml) werestirred and heated overnight at 64° C. Most of the solvent wasevaporated under vacuum and the residue dissolved in water. Sodiumbicarbonate solution (5% w/w, 5 ml) was added and the solution dialysedthree times with water. This solution was evaporated to dryness and theresidue dissolved in sodium bicarbonate solution (10 ml), dialysed andevaporated as before. This process was repeated, the resulting solid wasdissolved in a small volume of water and the product precipitated withmethanol. This was dissolved in water and evaporated to dryness givingthe title compound (1.18 g).

¹H NMR (D₂O): δ 3.9 (m, 24H), 3.2 (m, 24H), 3.55-3.65 (m, 16H), 3.9 (m,8H), 4.05 (m, 8H), 5.15 (s, 8H)

EXAMPLE 186-per-deoxy-6-per(2,2-di(hydroxymethyl)-3-hydroxy-propyl)thio-γ-cyclodextrin

Per-6-deoxy-per-6-thio-γ-cyclodextrin (500 mg; Example 17),3-bromo-2,2-dihydroxy-methylpropanol (670 mg), cesium carbonate (550 mg)and dimethylformamide (10 ml) were heated and stirred for 35 days at 65°C. until analysis by LCMS showed conversion to the required product. Themixture was evaporated to dryness, dissolved in water, dialysed againstwater, evaporated to low volume and precipitated with acetone. Dryingunder vacuum gave the title compound (550 mg). Mass spec. FIA (M−H)2369.

¹H NMR (D₂O): δ 2.84 (m, 16H), 3.15 (m, 8H), 3.24 (m, 8H), 3.69 (s,64H), 3.85-4.19 (m, 16H), 5.25 (s, 8H).

EXAMPLE 196-per-deoxy-6-per(3-(tetrazol-5-yl)propyl)thio-γ-cyclodextrin, sodiumsalt

Per-6-deoxy-per-6-thio-γ-cyclodextrin (1 g), 4-bromobutyronitrile (1 g),cesium carbonate (1 g) and dimethylformamide (10 ml) were stirredtogether at 60° C. over the weekend. The mixture was cooled, water addedand the precipitate separated by centrifuge. After washing and dryingthe per-butyronitrile (1.4 g) was obtained. This product (1 g), sodiumazide (1.3 g), triethylamine hydrochloride (2.8 g) and dimethylformamide(13 ml) were stirred and heated together for 7 days at 100° C. Themixture was cooled, diluted with water, acidified and the precipitatedfiltered off. This was washed with water, sonicated with methanol,separated by centrifuge, dried and dissolved in sodium hydroxidesolution (M, 10 ml), filtered and dialysed to neutrality. This solutionwas evaporated to dryness to give the title compound (600 mg). Massspec. (M−2H) 1152.8.

¹H NMR (D₂O); δ 1.95 (m, 16H), 2.55 (m, 16H), 2.85 (m, 24H), 3.05 (d,8H), 3.5 (m, 8H), 3.6 (m, 8H), 3.9 (m, 16H), 5.06 (s, 8H).

EXAMPLE 20

Reversal of Neuromuscular Blockade in Anaesthetized Guinea Pigs in vivo.

Male Dunkin-Hartley guinea pigs (bodyweight: 600-900 g) wereanaesthetized by i.p. administration of 10 mg/kg pentobarbitone and 1000mg/kg urethane. After tracheotomy, the animals were artificiallyventilated using a Harvard small animal ventilator. A catheter wasplaced into the carotid artery for continuous monitoring of arterialblood pressure and the taking of blood samples for blood gas analysis.Heart rate was derived from the blood pressure signal. The sciatic nervewas stimulated (rectangular pulses of 0.5 ms duration at 10 s (0.1 Hz)intervals at a supramaximal voltage, using a Grass S88 Stimulator) andthe force of M. gastrocnemius contractions was measured using a GrassFT03 force-displacement transducer. Contractions, blood pressure andheart rate were recorded on a multichannel Grass 7D recorder. Catheterswere placed in both jugular veins. One catheter was used for thecontinuous infusion of a neuromuscular blocking agent. The infusion rateof the neuromuscular blocking agent was increased until a steady-stateblock of 85-90% was obtained. The other catheter was used foradministration of increasing doses of the reversal agent. Duringcontinuous infusion of the neuromuscular blocking agent, single doses ofincreasing concentration of reversal agent were given. At the end of theexperiment, the measured force of muscle contractions was plottedagainst the concentration of reversal agent, and using regressionanalysis techniques, the 50% reversal concentration was calculated,Results for the reversal of the neuromuscular block, induced by themuscle relaxant rocuronium bromide (Roc), by the 6-mercapto-cyclodextrinderivatives of Examples 1-19 are presented in Table 1. For comparison,the reversal activity of the parent compounds β-cyclodextrin andγ-cyclodextrin are included as well.

TABLE I Dose (ED₅₀, μmol.kg⁻¹) producing 50% reversal of steady-stateneuromuscular block in anaesthetized guinea pigs and concentration atmamimum reversal. % max reversal ED₅₀ at conc. Compound μmol.kg⁻¹(μmol.kg⁻¹) γ-cyclodextrin (γ-CD) 4 104 (47) β-cyclodextrin (β-CD) 20 93(113) 6-mono-deoxy-6-mono-(4-carboxy- 0.94 102 (8.0) phenyl)thio-γ-cyclodextrin, Na salt (example 1)6-mono-deoxy-6-mono-(2-carboxyphenyl) 1.30 93 (11) thio-γ-cyclodextrin(example 2) 6-per-deoxy-6-per-(3-carboxyphenyl)thio-γ- 0.28 102 (1.28)cyclodextrin (example 3) 6-per-deoxy-6-per-(2-carboxyethyl)thio-γ- 0.0997 (0.53) cyclodextrin, Na salt (example 4)6-per-deoxy-6-per-(5-carboxypentyl)thio-γ- 0.74 78 (2.5) cyclodextrin,Na salt (example 5) 6-per-deoxy-6-per-(3-carboxypropyl)thio-γ- 0.09 108(0.48) cyclodextrin, Na salt (example 6)6-per-deoxy-6-per-carboxymethylthio-γ- 0.21 88 (1.92) cyclodextrin, Nasalt (example 7) 6-per-deoxy-6-per-(4-carboxyphenyl)thio-γ- 0.10 95(0.48) cyclodextrin, Na salt (example 8)6-per-deoxy-6-per-(4-carboxyphenylmethyl) 0.13 100 (0.50)thio-γ-cyclodextrin, Na salt (example 9)6-per-deoxy-6-per-(3-amidopropyl)thio-γ- 0.57 94 (33) cyclodextrin(example 10) 6-per-deoxy-6-per-(5-hydroxy-3-oxa-pentyl) 0.47 92 (2.1)thio-γ-cyclodextrin (example 11) 6-per-deoxy-6-per-[(2(2-carboxybenzoyl)0.085 95 (0.48) amino)ethyl]-thio-γ-cyclodextrin, sodium salt (example12) 6-per-deoxy-6-per-(2-hydroxyethyl)thio-γ- 0.20 96 (2.0) cyclodextrin(example 13) 6-per-deoxy-6-per-(N-methylamidomethyl) 1.54 102 (7.3)thio-γ-cyclodextrin (example 14)6-per-deoxy-6-per-(2-carboxypropyl)thio-γ- 0.10 103 (0.48) cyclodextrin,sodium salt. (example 15) 6-per-deoxy-6-per-(3-carboxypropyl)thio-β- 0.5100 (3.2) cyclodextrin, sodium salt (example 16)6-per-deoxy-6-per-(2-sulfoethyl)thio-γ- 0.055 106 (1.7) cyclodextrin,sodium salt (example 17) 6-per-deoxy-5-per-(2,2-di(hydroxymethyl)- 2.963 (4.9) 3-hydroxy-propyl)thio-y-cyclodextrin (example 18)6-per-deoxy-6-per-(3-(tetrazol-5-yl)propyl) 0.22 109 (1.02)thio-γ-cyclodextrin, sodium salt (example 19)

1. A kit for providing neuromuscular block and its reversal comprising(a) an aminosteroidal neuromuscular blocking agent and (b) a reversalagent selected from the group consisting of6-per-deoxy-6-per-(2-carboxyethyl)thio-γ-cyclodextrin;6-per-deoxy-6-per-(3-carboxypropyl)thio-γ-cyclodextrin;6-per-deoxy-6-per-(4-carboxyphenyl)thio-γ-cyclodextrin;6-per-deoxy-6-per-(4-carboxyphenylmethyl)thio-γ-cyclodextrin;6-per-deoxy-6-per-(2-carboxypropyl)thio-γ-cyclodextrin; and6-per-deoxy-6-per-(2-sulfoethyl)thio-γ-cyclodextrin; or apharmaceutically acceptable salt thereof.
 2. A kit according to claim 1wherein the aminosteroidal neuromuscular blocking agent is selected fromthe group consisting of rocuronium bromide, vecuronium bromide,pancuronium bromide, and rapacuronium bromide, or anotherpharmaceutically acceptable salt thereof.
 3. A kit according to claim 2,wherein the neuromuscular blocking agent is rocuronium bromide and thereversal agent is 6-per-deoxy-6per-(2-carboxyethyl)thio-γ-cyclodextrinsodium salt.
 4. A method of treatment of a patient with a neuromuscularblocking agent comprising the steps or (a) inducing the neuromuscularblock by administration of an effective amount of an aminosteroidalblocking agent selected from the group consisting of rocuronium bromide,vecuronium bromide, pancuronium bromide and rapacuronium bromide, oranother pharmaceutically acceptable salt thereof, and (b) reversing thedrug-induced neuromuscular block by administration of an effectiveamount of at least one reversal agent selected from the group consistingof 6-per-deoxy-6-per-(2-carboxyethyl)thio-γ-cyclodextrin;6-per-deoxy-6-per-(3-carboxypropyl)thio-γ-cyclodextrin;6-per-deoxy-6-per-(4-carboxyphenyl)thio-γ-cyclodextrin;6-per-deoxy-6-per-(4-carboxyphenylmethyl)thio-γ-cyclodextrin;6-per-deoxy-6-per-(2-carboxypropyl)thio-γ-cyclodextrin; and6-per-deoxy-6-per-(2-sulfoethyl)thio-γ-cyclodextrin; or apharmaceutically acceptable salt thereof.
 5. The method of treatmentaccording to claim 4, wherein the neuro-muscular blocking agent isrocuronium bromide and the reversal agent is6-per-deoxy-6-per-(2-carboxyethyl)thio-γ-cyclodextrin sodium salt.